microRNA expression profile in porcine oocytes with different developmental competence derived from large or small follicles

Abstract

Oocyte developmental competence is acquired during folliculogenesis and regulated by complex molecular mechanisms. Several molecules are involved in these mechanisms, including microRNAs (miRNAs) that are essential for oocyte‐specific processes throughout the development. The objective of this study was to identify the expression profile of miRNAs in porcine oocytes derived from follicles of different sizes using RNA deep sequencing. Oocytes were aspirated from large (LO; 3–6 mm) or small (SO; 1.5–1.9 mm) follicles and tested for developmental competence and chromatin configurations. Small RNA libraries were constructed from both groups and then sequenced in an Illumina NextSeq. 500. Oocytes from the LO group exhibited higher developmental competence and different chromatin configuration compared with oocytes from the SO group. In total, 167 and 162 known miRNAs were detected in the LO and SO groups, respectively. MiR‐205, miR‐16, miR‐148a‐3p, and miR‐125b were among the top 10 highly expressed miRNAs in both groups. Eight miRNAs were differentially expressed (DE) between both groups. Target gene prediction and pathway analysis revealed 46 pathways that were enriched with miRNA‐target genes. The oocyte meiosis pathway and signaling pathways including FoxO, PI3K‐Akt, and cAMP were predictably targeted by DE miRNAs. These results give more insights into the potential role of miRNAs in regulating the oocyte development.

Global transcriptome analysis of porcine oocytes in correlation with follicle size

“…The interaction networking of DEGs commonly involved in highly enriched biological processes including extracellular matrix organization, insulin‐like growth factor binding, microtubule cytoskeleton reorganization, and cell junction organization is presented in Figure . Comparison analysis with our previously published data (Gad et al, ) showed that a list of 99 DEGs between the LO and SO group was the same as the predicted target genes of the DE miRNAs in the same groups (Table S5). These genes are involved in different signaling pathways, gene expression regulation, and functions related to an extracellular matrix organization.…”

Research on miRNAs of exosomes’ role on porcine oocyte development by extracted from follicular fluid

“…Based on the above research results, these predicted ten relevant miRNAs among these four groups (miR-125b, let-7d-5p,miR-200b, miR-26a, miR-92a, miR-221-3p, miR-21-5p, miR-141,miR-99a-5p and miR-10a-5p as shown in Figure 1-6) related to porcine oocyte maturation are choose to take Miranda and RNA software to predict the targeting regulation relationship between the whole species of miRNAs (taking the intersection results of the two prediction software as the final target gene prediction results). The miR-125b, let-7d-5p and miR-200b were among the top 10 highly expressed miRNAs in four groups, which it is proved that it has potential role of miRNAs in regulating the oocyte development 17,18 . The parameters of this analysis are set as follows: energy_miranda<-20, score_miranda>150, energy_RNAhybird <-25.…”

Inhibition of miR-152 During in Vitro Maturation Enhances the Developmental Potential of Porcine Embryos

“…In our previous study, we compared the miRNA expression pro le of the growing and fully grown porcine oocytes based on the follicle size using small RNA high-throughput sequencing technology [16]. Results showed that miR-152 was among the signi cantly upregulated miRNAs in the large fully grown oocytes compared to the small growing ones.…”

“…The reaction mixture was incubated at 16 °C, then 42 °C for 30 min., followed by 85 °C for 5 min. A QuantaLife QX200 ddPCR system (Bio-Rad Inc.) was used for miRNA quanti cation as previously described [16]. The expression of miR-152 was normalized to miR-125b and miR-26a, as both exhibited high stability in porcine oocytes [16].…”

“…A QuantaLife QX200 ddPCR system (Bio-Rad Inc.) was used for miRNA quanti cation as previously described [16]. The expression of miR-152 was normalized to miR-125b and miR-26a, as both exhibited high stability in porcine oocytes [16]. In addition, miR-26a has been previously reported to be the most stable expressed miRNA in porcine oocytes [19].…”